Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 2001 May 15;356(Pt 1):53–60. doi: 10.1042/0264-6021:3560053

Cloning of a lymphatic peptide/histidine transporter.

K Sakata 1, T Yamashita 1, M Maeda 1, Y Moriyama 1, S Shimada 1, M Tohyama 1
PMCID: PMC1221811  PMID: 11336635

Abstract

Although peptide transport across the plasma membrane has been characterized well in the kidney and the intestine, the functional relevance of this transport in other organs has not been addressed. Here we report the cloning of a cDNA for a novel peptide/histidine transporter found in the rat (rPHT2), whose mRNA is expressed mainly in the lymphatic system. rPHT2 encodes a protein of 582 amino acids and showed 49% identity with the brain PHT (PHT1) [Yamashita, Shimada, Guo, Sato, Kohmura, Hayakawa, Takagi and Tohyama (1997) J. Biol. Chem. 272, 10205-10211]. rPHT2 mRNA was abundant in lung, spleen and thymus, and detected faintly in brain, liver, adrenal gland and heart by Northern-blot analysis and reverse transcriptase PCR (RT-PCR). Intense signals for the gene were found in immunocytes using in situ hybridization. Ectopic expression of rPHT2 protein in HEK-293T cells and BHK cells was not found on the cell surface, but was found on the lysosomal membrane using light- and electron-microscopic analysis. Recombinant rPHT2 protein reconstituted into liposomes showed proton-dependent transport activity with histidine and histidyl-leucine. These findings suggest that rPHT2 is involved in the protein catabolic pathway in the lymphatic system.

Full Text

The Full Text of this article is available as a PDF (353.4 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bockman D. E., Ganapathy V., Oblak T. G., Leibach F. H. Localization of peptide transporter in nuclei and lysosomes of the pancreas. Int J Pancreatol. 1997 Dec;22(3):221–225. doi: 10.1007/BF02788388. [DOI] [PubMed] [Google Scholar]
  2. Chen J. W., Murphy T. L., Willingham M. C., Pastan I., August J. T. Identification of two lysosomal membrane glycoproteins. J Cell Biol. 1985 Jul;101(1):85–95. doi: 10.1083/jcb.101.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cooper P. R., Nowak N. J., Higgins M. J., Church D. M., Shows T. B. Transcript mapping of the human chromosome 11q12-q13.1 gene-rich region identifies several newly described conserved genes. Genomics. 1998 May 1;49(3):419–429. doi: 10.1006/geno.1998.5291. [DOI] [PubMed] [Google Scholar]
  4. Cooper P. R., Nowak N. J., Higgins M. J., Simpson S. A., Marquardt A., Stoehr H., Weber B. H., Gerhard D. S., de Jong P. J., Shows T. B. A sequence-ready high-resolution physical map of the best macular dystrophy gene region in 11q12-q13. Genomics. 1997 Apr 15;41(2):185–192. doi: 10.1006/geno.1997.4660. [DOI] [PubMed] [Google Scholar]
  5. Cowles C. R., Odorizzi G., Payne G. S., Emr S. D. The AP-3 adaptor complex is essential for cargo-selective transport to the yeast vacuole. Cell. 1997 Oct 3;91(1):109–118. doi: 10.1016/s0092-8674(01)80013-1. [DOI] [PubMed] [Google Scholar]
  6. Döhner H., Stilgenbauer S., James M. R., Benner A., Weilguni T., Bentz M., Fischer K., Hunstein W., Lichter P. 11q deletions identify a new subset of B-cell chronic lymphocytic leukemia characterized by extensive nodal involvement and inferior prognosis. Blood. 1997 Apr 1;89(7):2516–2522. [PubMed] [Google Scholar]
  7. Fei Y. J., Ganapathy V., Leibach F. H. Molecular and structural features of the proton-coupled oligopeptide transporter superfamily. Prog Nucleic Acid Res Mol Biol. 1998;58:239–261. doi: 10.1016/s0079-6603(08)60038-0. [DOI] [PubMed] [Google Scholar]
  8. Fei Y. J., Kanai Y., Nussberger S., Ganapathy V., Leibach F. H., Romero M. F., Singh S. K., Boron W. F., Hediger M. A. Expression cloning of a mammalian proton-coupled oligopeptide transporter. Nature. 1994 Apr 7;368(6471):563–566. doi: 10.1038/368563a0. [DOI] [PubMed] [Google Scholar]
  9. Frommer W. B., Hummel S., Rentsch D. Cloning of an Arabidopsis histidine transporting protein related to nitrate and peptide transporters. FEBS Lett. 1994 Jun 27;347(2-3):185–189. doi: 10.1016/0014-5793(94)00533-8. [DOI] [PubMed] [Google Scholar]
  10. Ganapathy V., Pashley D. H., Fonteles M. C., Leibach F. H. Peptidases and peptide transport in mammalian kidney. Contrib Nephrol. 1984;42:10–18. doi: 10.1159/000409956. [DOI] [PubMed] [Google Scholar]
  11. Höning S., Sandoval I. V., von Figura K. A di-leucine-based motif in the cytoplasmic tail of LIMP-II and tyrosinase mediates selective binding of AP-3. EMBO J. 1998 Aug 10;17(5):1304–1314. doi: 10.1093/emboj/17.5.1304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Isenman L. D., Dice J. F. Selective release of peptides from lysosomes. J Biol Chem. 1993 Nov 15;268(32):23856–23859. [PubMed] [Google Scholar]
  13. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  14. Lewis V., Green S. A., Marsh M., Vihko P., Helenius A., Mellman I. Glycoproteins of the lysosomal membrane. J Cell Biol. 1985 Jun;100(6):1839–1847. doi: 10.1083/jcb.100.6.1839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Liu W., Liang R., Ramamoorthy S., Fei Y. J., Ganapathy M. E., Hediger M. A., Ganapathy V., Leibach F. H. Molecular cloning of PEPT 2, a new member of the H+/peptide cotransporter family, from human kidney. Biochim Biophys Acta. 1995 May 4;1235(2):461–466. doi: 10.1016/0005-2736(95)80036-f. [DOI] [PubMed] [Google Scholar]
  16. Miyamoto K., Shiraga T., Morita K., Yamamoto H., Haga H., Taketani Y., Tamai I., Sai Y., Tsuji A., Takeda E. Sequence, tissue distribution and developmental changes in rat intestinal oligopeptide transporter. Biochim Biophys Acta. 1996 Feb 7;1305(1-2):34–38. doi: 10.1016/0167-4781(95)00208-1. [DOI] [PubMed] [Google Scholar]
  17. Moriyama Y., Iwamoto A., Hanada H., Maeda M., Futai M. One-step purification of Escherichia coli H(+)-ATPase (F0F1) and its reconstitution into liposomes with neurotransmitter transporters. J Biol Chem. 1991 Nov 25;266(33):22141–22146. [PubMed] [Google Scholar]
  18. Niwa H., Yamamura K., Miyazaki J. Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene. 1991 Dec 15;108(2):193–199. doi: 10.1016/0378-1119(91)90434-d. [DOI] [PubMed] [Google Scholar]
  19. Ogihara H., Saito H., Shin B. C., Terado T., Takenoshita S., Nagamachi Y., Inui K., Takata K. Immuno-localization of H+/peptide cotransporter in rat digestive tract. Biochem Biophys Res Commun. 1996 Mar 27;220(3):848–852. doi: 10.1006/bbrc.1996.0493. [DOI] [PubMed] [Google Scholar]
  20. Rentsch D., Laloi M., Rouhara I., Schmelzer E., Delrot S., Frommer W. B. NTR1 encodes a high affinity oligopeptide transporter in Arabidopsis. FEBS Lett. 1995 Aug 21;370(3):264–268. doi: 10.1016/0014-5793(95)00853-2. [DOI] [PubMed] [Google Scholar]
  21. Saito H., Okuda M., Terada T., Sasaki S., Inui K. Cloning and characterization of a rat H+/peptide cotransporter mediating absorption of beta-lactam antibiotics in the intestine and kidney. J Pharmacol Exp Ther. 1995 Dec;275(3):1631–1637. [PubMed] [Google Scholar]
  22. Saito H., Terada T., Okuda M., Sasaki S., Inui K. Molecular cloning and tissue distribution of rat peptide transporter PEPT2. Biochim Biophys Acta. 1996 Apr 26;1280(2):173–177. doi: 10.1016/0005-2736(96)00024-7. [DOI] [PubMed] [Google Scholar]
  23. Sakata K., Shimada S., Yamashita T., Inoue K., Tohyama M. Cloning of a bovine orphan transporter and its short splicing variant. FEBS Lett. 1999 Jan 29;443(3):267–270. doi: 10.1016/s0014-5793(98)01733-5. [DOI] [PubMed] [Google Scholar]
  24. Sandoval I. V., Arredondo J. J., Alcalde J., Gonzalez Noriega A., Vandekerckhove J., Jimenez M. A., Rico M. The residues Leu(Ile)475-Ile(Leu, Val, Ala)476, contained in the extended carboxyl cytoplasmic tail, are critical for targeting of the resident lysosomal membrane protein LIMP II to lysosomes. J Biol Chem. 1994 Mar 4;269(9):6622–6631. [PubMed] [Google Scholar]
  25. Sandoval I. V., Bakke O. Targeting of membrane proteins to endosomes and lysosomes. Trends Cell Biol. 1994 Aug;4(8):292–297. doi: 10.1016/0962-8924(94)90220-8. [DOI] [PubMed] [Google Scholar]
  26. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sembries S., Pahl H., Stilgenbauer S., Döhner H., Schriever F. Reduced expression of adhesion molecules and cell signaling receptors by chronic lymphocytic leukemia cells with 11q deletion. Blood. 1999 Jan 15;93(2):624–631. [PubMed] [Google Scholar]
  28. Steiner H. Y., Song W., Zhang L., Naider F., Becker J. M., Stacey G. An Arabidopsis peptide transporter is a member of a new class of membrane transport proteins. Plant Cell. 1994 Sep;6(9):1289–1299. doi: 10.1105/tpc.6.9.1289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stöhr H., Marquardt A., Rivera A., Cooper P. R., Nowak N. J., Shows T. B., Gerhard D. S., Weber B. H. A gene map of the Best's vitelliform macular dystrophy region in chromosome 11q12-q13.1. Genome Res. 1998 Jan;8(1):48–56. doi: 10.1101/gr.8.1.48. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Takahashi Y., Miyata M., Zheng P., Imazato T., Horwitz A., Smith J. D. Identification of cAMP analogue inducible genes in RAW264 macrophages. Biochim Biophys Acta. 2000 Jul 24;1492(2-3):385–394. doi: 10.1016/s0167-4781(00)00133-0. [DOI] [PubMed] [Google Scholar]
  31. Thamotharan M., Lombardo Y. B., Bawani S. Z., Adibi S. A. An active mechanism for completion of the final stage of protein degradation in the liver, lysosomal transport of dipeptides. J Biol Chem. 1997 May 2;272(18):11786–11790. doi: 10.1074/jbc.272.18.11786. [DOI] [PubMed] [Google Scholar]
  32. Vega M. A., Rodriguez F., Seguí B., Calés C., Alcalde J., Sandoval I. V. Targeting of lysosomal integral membrane protein LIMP II. The tyrosine-lacking carboxyl cytoplasmic tail of LIMP II is sufficient for direct targeting to lysosomes. J Biol Chem. 1991 Sep 5;266(25):16269–16272. [PubMed] [Google Scholar]
  33. Vega M. A., Seguí-Real B., García J. A., Calés C., Rodríguez F., Vanderkerckhove J., Sandoval I. V. Cloning, sequencing, and expression of a cDNA encoding rat LIMP II, a novel 74-kDa lysosomal membrane protein related to the surface adhesion protein CD36. J Biol Chem. 1991 Sep 5;266(25):16818–16824. [PubMed] [Google Scholar]
  34. Yamashita T., Shimada S., Guo W., Sato K., Kohmura E., Hayakawa T., Takagi T., Tohyama M. Cloning and functional expression of a brain peptide/histidine transporter. J Biol Chem. 1997 Apr 11;272(15):10205–10211. doi: 10.1074/jbc.272.15.10205. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES